Characterisation of bound residues of [3H]triforine in the straw of barley grown in the field

Barley was grown in an experimental field and, at the growth stage J (during the stem extension stage when the second node of the stem was formed and the next-to-last leaf just visible), the aerial part of the plants was sprayed with an aqueous emulsion of a mixture of triforine and [³H]triforine (uniformly labelled in the piperazine ring) at the recommended rate of 250 g triforine ha^?1. Barley was harvested when ripe, and the straw was analysed separately. Extraction of the straw with methanol left methanol-insoluble solids containing an amount of radioactivity (the bound residue) which represented 88 % of the tritium (as with all the subsequently quoted percentages, relative to the total tritium incorporated into the straw). Methanol acidified with hydrochloric acid extracted a further 8% of the triforine-derived bound residues as radioactive iminodiacetic acid (0.4%). glycine (3.2%), serine (2.0%), ethanolamine (0.2%) and unidentified compounds (2.2%); a neutral detergent solution extracted a further 3% of the radioactive triforine-derived bound residues as unidentified compounds; these radioactive compounds, which were dissolved by the acidified methanol and by the neutral detergent solution, were thus complexed in the straw to straw constituents. An acid detergent solution extracted a further 58% of the total tritium, which was incorporated by means of σ chemical bonds into the hemicelluloses fraction. Acid hydrolysis of the hemicelluloses yielded a mixture of mono-saccharides; these were converted into a mixture of osazones (50%), recrystallised several times, and had a constant specific radioactivity, indicating that the mono-saccharides were tritiated. The plant solids (which contained 19% of the total tritium), left by the acid-detergent extraction, were processed and separated into the tritium-containing cellulose (13%) and lignin (6%) fractions. No piperazine was observed in the bound residue of straw. Biochemical pathways are suggested for the radioactive metabolites of [³H]triforine.     

Short communication: Satellite DNA-based species-specific identification of single individuals of the pinewood nematode <Emphasis Type="Italic">Bursaphelenchus xylophilus</Emphasis> (Nematoda: Aphelenchoididae)

The pinewood nematode Bursaphelenchus xylophilus is a severe pest of coniferous trees, and has been designated as a quarantine organism in the European Union. From the sequence of a satellite DNA family characterized in the genome of this nematode, we developed a PCR procedure that allowed the specific discrimination of this species from closely related Bursaphelenchus species found on coniferous trees. Moreover, because of the repetitive nature of satellite DNA, positive amplification was achieved from B. xylophilus single individuals, which should contribute to an easy diagnostic procedure for assisting in the management of this major pest of conifer forests.     

Durably controlling bovine hypodermosis

Cattle hypodermosis, due to insect larvae, is widely spread over the northern hemisphere. Very efficient insecticides are available and their use in most countries are done on an individual level but never cover the whole cattle population of a country. Untreated animals remain the reservoir of the disease and annually re-infest the cattle population. The economic effects of this disease on animal production (meat, milk and the leather industry) but also on the general cattle health status, have led many European countries to launch organised control programs. The first example of definitive hypodermosis control goes back one hundred years ago when Danish farmers eradicated hypodermosis from the Danish islands by manual elimination of the warbles. Since then, more and more European countries have considered the feasibility and economic returns of such programs. The various factors which foster these programs are related to (i) biological factors, (parasite cattle specificity, synchronous biological cycles of both species of insects involved), (ii) the development of more and more efficient insecticides used only once a year by systemic application, with high efficiency at very low dosages against the first larval stage of Hypoderma spp., (iii) the development of acute techniques of detection of the disease for the monitoring of hypodermosis free countries and (iv) the durable successful results obtained in more and more European countries. Although the programs were imposed by different partners of the livestock channel production (farmers, dairy industry, leather industry) and have been engaged within the last 50 years in many European countries (Denmark, The Netherlands, Ireland, the United Kingdom, the Czech Republic, Germany, France and Switzerland) common features have emerged among these different eradication programs. They all need a preliminary statement of the economic impact of this pest and the farmers' awareness of the economic returns of such programs. The programs' efficacy depends: (i) on a good knowledge ofthe epidemiology of the parasites, (ii) on the simultaneous implementation of the control program on the whole national cattle population whatever the structure monitoring the treatments (veterinary services, farmers association), (iii) on a national Warble fly legislation making the treatments compulsory and (iv) on an acute epidemiological survey as soon as the status of a hypodermosis free country is reached and the treatments are suspended. The sanitary and financial returns of such programs are a benefit to all the partners of livestock production, to the quality of the environment and to the consumers.     

Use of a serum-free medium to produce in vitro Neospora caninum and Toxoplasma gondii tachyzoites on Vero cells

Neospora caninum and Toxoplasma gondii are cyst-forming coccidian parasites of human and veterinary clinical relevance. In vitro cultivation of the protozoans using Vero cells is usually performed in order to produce antigenic materials. Quantitative and qualitative comparisons of Vero cells grown in RPMI medium supplemented either with foetal calf serum (FCS), horse serum (HS) or a specific serum-free additive (DefCell) were performed. A serum-free cell culture system used to propagate N. caninum (NC-1 isolate) and T. gondii tachyzoites (Rh stain) were compared with the other two cell culture systems. FCS supplemented media was found to be more effective than the others in promoting Vero cells and N. caninum tachyzoites. However, it was found unable to support adequate T. gondii tachyzoite proliferation. Vero cells, T. gondii and N. caninum tachyzoite production gave similar growth patterns with either HS or DefCell supplemented media. Defcell was considered as a good alternative to supplement culture medium.     

Implication of Systemic Induced Resistance in the Suppression of Fusarium Wilt of Tomato by Pseudomonas fluorescens WCS417r and by Nonpathogenic Fusarium oxysporum Fo47

Fluorescent pseudomonads and nonpathogenic Fusarium oxysporum have been shown to suppress fusarium wilts. This suppression has been related to both microbial antagonism and induced resistance.The aim of the present study was to assess the relative importance of systemic induced resistance in the suppression of fusarium wilt of tomato in commercial-like conditions by a reference strain of each type of microorganism (P. fluorescens WCS417r and nonpathogenic F. oxysporum Fo47). The spatial separation of the pathogen and the biocontrol strains excluded any possible microbial antagonism and implicated the involvement of the systemic induced resistance; whereas the absence of any separation between these microorganisms allowed the expression of both mechanisms. Since systemic induced resistance has often been associated with the synthesis of PR-proteins, their accumulation in tomato plants inoculated with WCS417r or with Fo47 was determined.The analysis of the results indicates that the suppression of fusarium wilt by P. fluorescens WCS417r was ascribed to systemic induced resistance without any detection of the PR-proteins tested (PR-1 and chitinases). In contrast, the suppression achieved by nonpathogenic F. oxysporum Fo47 appeared to be mainly ascribed to microbial antagonism but also to a lesser extent to systemic induced resistance. This induced resistance could be related to the accumulation of PR-1 and chitinases.The possible relationship between the ability of Fo47 to suppress fusarium wilt more efficiently than WCS417r and its ability to show both mechanisms is discussed.     

Effect of soil nitrogen supply on carbon assimilation by tree stems

• ? Nitrogen (N) is one of the most important resources for plants, generally enhancing leaf photosynthesis because a large part of it is allocated to Rubisco and thylakoïds. This is well known in leaves where photosynthesis (i.e. gas exchange, Rubisco activity, chlorophyll content) is positively correlated to leaf N content.
• ? In order to test this hypothesis in stems, N concentration, CO₂ exchange and also Rubisco and PEP carboxylase activities were measured in summer on current-year stems of young European beeches (Fagus sylvatica L.) growing on soils of different N content.
• ? The CO₂ refixation rate of stems increased from 58.5% to 74.3% when stem N concentration increased from 5.7 to 10.1 mg g^?1 DW. A hyperbolic relationship was obtained between stem gross photosynthesis and N concentration, with an x-intercept of 0.3 mmol N g^?1 DW. Stem PEP carboxylase activity was higher in stems than in leaves and increased with stem N concentration whereas Rubisco activity did not change between treatments in both tissues.
• ? In spite of a low nitrogen investment in stem photosynthesis (low PNUE), these results suggest that (1) stems invest more N in CO₂ refixation when more N is widely available, (2) stem photosynthesis is able to operate at low N concentration and (3) stem PEP carboxylase is involved in stem carbon refixation, but also simultaneously supplies carbon skeletons for N assimilation.

     

Biotransformation of s-Triazine Herbicides and Related Degradation Products in Liquid Cultures by the White Rot Fungus Phanerochaete chrysosporium

The ability of the white rot basidiomycete Phanerochaete chrysosporium to transform s-triazine herbicides has been investigated in laboratory experiments. The chlorinated metabolites formed during atrazine N-dealkylations were not further transformed by the fungus, whereas hydroxy-atrazine was converted to an unknown product. P. chrysosporium was also able to carry out the N-dealkylation of the herbicides simazine, propazine and terbuthylazine. Herbicide metabolism was not supported by purified peroxidases. The highest rates of herbicide N-dealkylation were obtained in liquid cultures maintained under moderate temperature allowing a long mycelium growing phase. Atrazine transformation was found to be supported by the mycelium, which contained significant amounts of microsomal cytochrome P450. Herbicide N-dealkylation was decreased in the presence of 1-aminobenzotriazole, in agreement with the involvement of P450 monooxygenases in atrazine metabolism. © 1997 SCI.     

Effect of vitamin A on the skeletal morphogenesis of European sea bass,Dicentrarchus labrax (Linnaeus, 1758)

The purpose of this study was to examine the effects of vitamin A levels in the larval diet of Dicentrarchus labrax, (a) on the shape of juvenile skeleton and (b) on the development of pelvic fins. To exclude the side‐effects of skeletal abnormalities on skeleton shape, morphometric analysis was performed on normal individuals only. Geometric morphometric analysis revealed a significant effect of larval nutrition on the shape of juvenile skeleton (Wilk's λ=0.0188, P<0.001), 55 days after the application of the different nutritional regimes. Between‐groups shape differences mainly concerned the position of anterior dorsal pterygiophore, which was associated with a significant (P<0.05) increase of the number of dorsal lepidotrichia and hard spines at low levels of vitamin A (3–9 × 10³ IU kg^?1 dry food). At the lowest vitamin A levels (3 × 10³ IU kg^?1 dry food), the development of pelvic fins was abnormal, with severe bilateral (44±10%, mean±SD) or unilateral (22±9%) lack of the fin lepidotrichia, hard spines and their supporting elements. In the 82–94% of the unilaterally deformed individuals, deformation concerned the left pelvic fin. Concerning the shape of pelvic fins in the normally developed individuals, results demonstrated a significant effect of nutritional conditions on the size and position of all the elements of the pelvic fin area (basipterygium, lower metacleithrum, cleithrum, scapula) (Wilk's λ=0.3087, P<0.001). Our results demonstrate a significant nutritionally driven phenotypic plasticity in D. labrax, with dietary levels of vitamin A determining the skeleton shape and the left–right symmetry of pelvic fins.